The promoter of the leghaemoglobin gene VfLb29: Functional analysis and identification of modules necessary for its activation in the infected cells of root nodules and in the arbuscule-containing cells of mycorrhizal roots

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Original languageEnglish
Pages (from-to)799-806
Number of pages8
JournalJournal of experimental botany
Volume56
Issue number413
Publication statusPublished - 1 Mar 2005
Externally publishedYes

Abstract

In this study the further characterization of the Vicia faba leghaemoglobin promoter pVfLb29 is presented that was previously shown to be specifically active in the infected cells of root nodules and in arbuscule-containing cells of mycorrhizal roots. Using promoter studies in transgenic hairy roots of the Pisum sativum mutant RisNod24, disabled in the formation of functional arbuscules, VfLb29 promoter activity is assigned to later stages of arbuscule development. In order to narrow down the regions containing cis-acting elements of pVfLb29, the activity of five VfLb29 promoter deletions (-797/-31 to -175/-31 in relation to the start codon) fused to the gusAint coding region were tested in transgenic V. hirsuta hairy roots. The results specify a promoter region ranging from position -410 to -326 (85 bp) as necessary for gus expression in arbuscule-containing cells, whereas this segment is not involved in the nodule-specific activity. Sequence analysis of the pVfLb29 fragment -410/-326 (85 bp) revealed sequence motifs previously shown to be cis-acting elements of diverse promoters. To investigate the autonomous function of pVfLb29 regions for activation in arbuscule-containing cells, different regions of pVfLb29 from positions -410 to -198 were used to prepare chimeric promoter constructs for trans-activation studies. These fragments alone did not activate the mycorrhiza inactive promoter of the Vicia faba leghaemoglobin gene VfLb3, showing that the activation of pVfLb29 in arbuscule-containing cells is governed by a complex regulatory system that requires at least two modules located between position -410 and -31 of the VfLb29 gene.

Keywords

    Arbuscules, Leghaemoglobin promoter, Mycorrhizal roots, Root nodules, Vicia faba

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The promoter of the leghaemoglobin gene VfLb29: Functional analysis and identification of modules necessary for its activation in the infected cells of root nodules and in the arbuscule-containing cells of mycorrhizal roots. / Fehlberg, Vera; Vieweg, Martin F.; Dohmann, Esther M.N. et al.
In: Journal of experimental botany, Vol. 56, No. 413, 01.03.2005, p. 799-806.

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title = "The promoter of the leghaemoglobin gene VfLb29: Functional analysis and identification of modules necessary for its activation in the infected cells of root nodules and in the arbuscule-containing cells of mycorrhizal roots",
abstract = "In this study the further characterization of the Vicia faba leghaemoglobin promoter pVfLb29 is presented that was previously shown to be specifically active in the infected cells of root nodules and in arbuscule-containing cells of mycorrhizal roots. Using promoter studies in transgenic hairy roots of the Pisum sativum mutant RisNod24, disabled in the formation of functional arbuscules, VfLb29 promoter activity is assigned to later stages of arbuscule development. In order to narrow down the regions containing cis-acting elements of pVfLb29, the activity of five VfLb29 promoter deletions (-797/-31 to -175/-31 in relation to the start codon) fused to the gusAint coding region were tested in transgenic V. hirsuta hairy roots. The results specify a promoter region ranging from position -410 to -326 (85 bp) as necessary for gus expression in arbuscule-containing cells, whereas this segment is not involved in the nodule-specific activity. Sequence analysis of the pVfLb29 fragment -410/-326 (85 bp) revealed sequence motifs previously shown to be cis-acting elements of diverse promoters. To investigate the autonomous function of pVfLb29 regions for activation in arbuscule-containing cells, different regions of pVfLb29 from positions -410 to -198 were used to prepare chimeric promoter constructs for trans-activation studies. These fragments alone did not activate the mycorrhiza inactive promoter of the Vicia faba leghaemoglobin gene VfLb3, showing that the activation of pVfLb29 in arbuscule-containing cells is governed by a complex regulatory system that requires at least two modules located between position -410 and -31 of the VfLb29 gene.",
keywords = "Arbuscules, Leghaemoglobin promoter, Mycorrhizal roots, Root nodules, Vicia faba",
author = "Vera Fehlberg and Vieweg, {Martin F.} and Dohmann, {Esther M.N.} and Natalija Hohnjec and Alfred P{\"u}hler and Perlick, {Andreas M.} and Helge K{\"u}ster",
note = "Funding information: We would like to thank the Deutsche Forschungsgemeinschaft for supporting this work in the frame of the SPP1084 (MolMyk) projects Pe814/1-2 and Ku1478/2-3. We are grateful to Gerard Duc (INRA Dijon, France) for providing seeds of the P. sativum mutant RisNod24 and to Rene Geurts (Wageningen University, Netherlands) for supplying the pRedRoot plasmid prior to publication.",
year = "2005",
month = mar,
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language = "English",
volume = "56",
pages = "799--806",
journal = "Journal of experimental botany",
issn = "0022-0957",
publisher = "Oxford University Press",
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TY - JOUR

T1 - The promoter of the leghaemoglobin gene VfLb29

T2 - Functional analysis and identification of modules necessary for its activation in the infected cells of root nodules and in the arbuscule-containing cells of mycorrhizal roots

AU - Fehlberg, Vera

AU - Vieweg, Martin F.

AU - Dohmann, Esther M.N.

AU - Hohnjec, Natalija

AU - Pühler, Alfred

AU - Perlick, Andreas M.

AU - Küster, Helge

N1 - Funding information: We would like to thank the Deutsche Forschungsgemeinschaft for supporting this work in the frame of the SPP1084 (MolMyk) projects Pe814/1-2 and Ku1478/2-3. We are grateful to Gerard Duc (INRA Dijon, France) for providing seeds of the P. sativum mutant RisNod24 and to Rene Geurts (Wageningen University, Netherlands) for supplying the pRedRoot plasmid prior to publication.

PY - 2005/3/1

Y1 - 2005/3/1

N2 - In this study the further characterization of the Vicia faba leghaemoglobin promoter pVfLb29 is presented that was previously shown to be specifically active in the infected cells of root nodules and in arbuscule-containing cells of mycorrhizal roots. Using promoter studies in transgenic hairy roots of the Pisum sativum mutant RisNod24, disabled in the formation of functional arbuscules, VfLb29 promoter activity is assigned to later stages of arbuscule development. In order to narrow down the regions containing cis-acting elements of pVfLb29, the activity of five VfLb29 promoter deletions (-797/-31 to -175/-31 in relation to the start codon) fused to the gusAint coding region were tested in transgenic V. hirsuta hairy roots. The results specify a promoter region ranging from position -410 to -326 (85 bp) as necessary for gus expression in arbuscule-containing cells, whereas this segment is not involved in the nodule-specific activity. Sequence analysis of the pVfLb29 fragment -410/-326 (85 bp) revealed sequence motifs previously shown to be cis-acting elements of diverse promoters. To investigate the autonomous function of pVfLb29 regions for activation in arbuscule-containing cells, different regions of pVfLb29 from positions -410 to -198 were used to prepare chimeric promoter constructs for trans-activation studies. These fragments alone did not activate the mycorrhiza inactive promoter of the Vicia faba leghaemoglobin gene VfLb3, showing that the activation of pVfLb29 in arbuscule-containing cells is governed by a complex regulatory system that requires at least two modules located between position -410 and -31 of the VfLb29 gene.

AB - In this study the further characterization of the Vicia faba leghaemoglobin promoter pVfLb29 is presented that was previously shown to be specifically active in the infected cells of root nodules and in arbuscule-containing cells of mycorrhizal roots. Using promoter studies in transgenic hairy roots of the Pisum sativum mutant RisNod24, disabled in the formation of functional arbuscules, VfLb29 promoter activity is assigned to later stages of arbuscule development. In order to narrow down the regions containing cis-acting elements of pVfLb29, the activity of five VfLb29 promoter deletions (-797/-31 to -175/-31 in relation to the start codon) fused to the gusAint coding region were tested in transgenic V. hirsuta hairy roots. The results specify a promoter region ranging from position -410 to -326 (85 bp) as necessary for gus expression in arbuscule-containing cells, whereas this segment is not involved in the nodule-specific activity. Sequence analysis of the pVfLb29 fragment -410/-326 (85 bp) revealed sequence motifs previously shown to be cis-acting elements of diverse promoters. To investigate the autonomous function of pVfLb29 regions for activation in arbuscule-containing cells, different regions of pVfLb29 from positions -410 to -198 were used to prepare chimeric promoter constructs for trans-activation studies. These fragments alone did not activate the mycorrhiza inactive promoter of the Vicia faba leghaemoglobin gene VfLb3, showing that the activation of pVfLb29 in arbuscule-containing cells is governed by a complex regulatory system that requires at least two modules located between position -410 and -31 of the VfLb29 gene.

KW - Arbuscules

KW - Leghaemoglobin promoter

KW - Mycorrhizal roots

KW - Root nodules

KW - Vicia faba

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U2 - 10.1093/jxb/eri074

DO - 10.1093/jxb/eri074

M3 - Article

C2 - 15668224

AN - SCOPUS:15944362788

VL - 56

SP - 799

EP - 806

JO - Journal of experimental botany

JF - Journal of experimental botany

SN - 0022-0957

IS - 413

ER -

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